CDH1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein
Frameshift = 95%.
Images
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein Frameshift = 95%
Alternative names
Arc 1, CADH1_HUMAN, CAM 120/80, CD 324, CD324 antigen, CDH1, CDHE, Cadherin-1, E-Cad/CTF3, E-cadherin, ECAD, Epithelial cadherin, LCAM, Liver cell adhesion molecule, UVO, Uvomorulin, cadherin 1 type 1 E-cadherin, epithelial calcium dependant adhesion protein
Recommended products
CDH1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein
Frameshift = 95%.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein Frameshift = 95%
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- CDH1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
E-Cadherin sometimes called CDH1 or Cadherin-1 is a protein that plays a role in cell-to-cell adhesion. This transmembrane protein has a molecular weight of approximately 120 kDa. E-Cadherin is mainly expressed in epithelial tissues of various organs including the skin and gut. Its adhesive function is made possible by its extracellular domain that facilitates homophilic binding between cells contributing to the maintenance of tissue architecture and cellular integrity.
- Biological function summary
E-Cadherin participates in establishing and maintaining adherens junctions which are vital for tissue structure. E-Cadherin operates as a core component of the cadherin-catenin complex which links the protein to the actin cytoskeleton. Through this linkage E-Cadherin plays an important role in signaling pathways that influence cellular growth and differentiation. The protein's ability to mediate intercellular connections also regulates cellular motility and supports basic aspects of cell behavior in epithelial tissues.
- Pathways
E-Cadherin influences both the Wnt signaling pathway and the epithelial-to-mesenchymal transition (EMT). Within the Wnt signaling pathway E-Cadherin partners with β-catenin a significant player in transcription regulation and cell signaling. Disruption in E-Cadherin's adhesive functionality can lead to increased β-catenin availability affecting downstream transcriptional control. In the EMT process E-Cadherin loss characterizes an important step in which cells gain migratory and invasive properties typically seen during metastasis in cancer progression.
- Associated diseases and disorders
E-Cadherin's role is prominent in cancer particularly in the context of gastric and breast cancers. Mutations or altered expression of E-Cadherin lead to diminished cell adhesion promoting tumorigenesis and metastatic spread. In gastric cancer for instance mutated E-Cadherin often associates with loss of epithelial function facilitating cancer cell dissemination. Additionally its loss or dysfunction may correlate with proteins such as β-catenin further impacting cancer progression and pathology.
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5 product images
Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/10000 dilution
Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free (Anti-E Cadherin antibody [EP700Y] - Low endotoxin, Azide free ab201499) at 1/10000 dilution
Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [EP700Y] - BSA and Azide free (Anti-E Cadherin antibody [EP700Y] - BSA and Azide free ab256580) at 1/10000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)
Lane 2: CDH1 knockout Raji cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: MDA-MB-231 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 105 kDa, 130 kDa
Next Generation Sequencing - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
1 bp insertion after Phe38 (allele 1), 4 bp deletion after Thr37 (allele 2), and 1 bp deletion after Thr37 (allele 3) of the WT protein
Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Western blot: Anti-CDH1 antibody [4A2] (Anti-E Cadherin antibody [4A2] ab231303) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-E Cadherin antibody [4A2] ab231303 was shown to bind specifically to CDH1. A band was observed at 130, 110, 55 kDa in wild-type A431 cell lysates with no signal observed at this size in CDH1 knockout cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-E Cadherin antibody [4A2] (Anti-E Cadherin antibody [4A2] ab231303) at 1 µg/mL
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lane 2: CDH1 knockout A431 cell lysate at 20 µg
Lane 3: Caco-2 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 97 kDa
Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Western blot: Anti-CDH1 antibody [EP700Y] (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772 was shown to bind specifically to CDH1. A band was observed at 130, 110, 80, 55, 40 kDa in wild-type A431 cell lysates with no signal observed at this size in CDH1 knockout cell line ab273747 (knockout cell lysate ab273781). To generate this image, wild-type and CDH1 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker (Anti-E Cadherin antibody [EP700Y] - Intercellular Junction Marker ab40772) at 1/1000 dilution
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lane 2: CDH1 knockout A431 cell lysate at 20 µg
Lane 3: Caco-2 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 97 kDa
Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [SP64] (Anti-E Cadherin antibody [SP64] ab227639) at 1/100 dilution
Lanes 1 - 4: Western blot - Anti-E Cadherin antibody [SP64] - BSA and Azide free (Anti-E Cadherin antibody [SP64] - BSA and Azide free ab240984) at 1/100 dilution
Lane 1: Wild-type A431 cell lysate at 20 µg
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell line (ab273747)
Lane 2: Western blot - Human CDH1 (E Cadherin) knockout A-431 cell lysate (ab273781)
Lane 2: CDH1 knockout A431 cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: MDA-MB-231 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 105 kDa, 130 kDa
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